NF kappaB p65 ELISA Kit is a single-wash 90-min Simplestep used to quantify NF kappaB p65 with a sensitivity of 5 µg/mL. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.
- Colorimetric Sandwich ELISA - 450 nm readout : works on any standard plate reader
- Cited in over 30 citations
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- International journal of molecular sciences 24:2023Bisphenol AF Induces Prostatic Dorsal Lobe Hyperplasia in Rats through Activation of the NF-κB Signaling Pathway.Applications:Unspecified applicationReactive species:Unspecified reactive speciesSisi Huang,Kaiyue Wang,Dongyan Huang,Xin Su,Rongfu Yang,Congcong Shao,Juan Jiang,Jianhui WuPubMed 38003411
- Heliyon 9:e220552023L. significantly alleviates cigarette smoke-induced acute lung injury by lowering NF-κB pathway activation and inflammatory mediators.Applications:Unspecified applicationReactive species:Unspecified reactive speciesNadia Hussain,Nadia Ikram,Kashif Ur Rehman Khan,Liaqat Hussain,Ali M Alqahtani,Taha Alqahtani,Musaddique Hussain,Muath Suliman,Mohammad Y Alshahrani,Basel SitohyPubMed 38045213
- Biomaterials research 27:652023Oxygen-loaded microbubble-mediated sonoperfusion and oxygenation for neuroprotection after ischemic stroke reperfusion.Applications:Unspecified applicationReactive species:Unspecified reactive speciesYi-Ju Ho,Hsiang-Lung Cheng,Lun-De Liao,Yu-Chun Lin,Hong-Chieh Tsai,Chih-Kuang YehPubMed 37415210
- Redox biology 63:1027142023LPS-induced mitochondrial dysfunction regulates innate immunity activation and α-synuclein oligomerization in Parkinson's disease.Applications:Unspecified applicationReactive species:Unspecified reactive speciesA Raquel Esteves,Diana F Silva,Diogo Banha,Emanuel Candeias,Beatriz Guedes,Sandra M CardosoPubMed 37120929
- Journal of physiology and pharmacology : an official journal of the Polish Physiological Society 73:20236-Shogaol protects against isoproterenol-induced cardiac injury in rats through attenutating oxidative stress, inflammation, apoptosis and activating nuclear respiratory factor-2/heme oxygenase-1 signaling pathway.Applications:Unspecified applicationReactive species:Unspecified reactive speciesH Li,J Shen,Y Zhang,L Hu,W LuoPubMed 37087565
- BioFactors (Oxford, England) 49:699-7112023Crocetin regulates Th1/Th2 and Th17/Treg balances, nitric oxide production, and nuclear localization of NF-κB in Th2-provoked and normal situations in human-isolated lymphocytes.Applications:Unspecified applicationReactive species:Unspecified reactive speciesVafa Baradaran Rahimi,Saeideh Saadat,Pouria Rahmanian Devin,Ali Jebalbarezy,Maryam Moqaddam,Mohammad Hossein Boskabady,Vahid Reza AskariPubMed 36747328
- Pharmaceuticals (Basel, Switzerland) 16:2023Ganetespib with Methotrexate Acts Synergistically to Impede NF-κB/p65 Signaling in Human Lung Cancer A549 Cells.Applications:Unspecified applicationReactive species:Unspecified reactive speciesGehad Subaiea,Syed Mohd Danish Rizvi,Hemant Kumar Singh Yadav,Turki Al Hagbani,Marwa Helmy Abdallah,El-Sayed Khafagy,Hosahalli Veerabhadrappa Gangadharappa,Talib Hussain,Amr Selim Abu LilaPubMed 37259378
- Molecules (Basel, Switzerland) 27:2022The Synergistic Hepatoprotective Activity of Rosemary Essential Oil and Curcumin: The Role of the MEK/ERK Pathway.Applications:Unspecified applicationReactive species:Unspecified reactive speciesMaged E Mohamed,Nancy S Younis,Hossam S El-Beltagi,Omar M MohafezPubMed 36558044
- Polymers 14:2022Preparation, Characterization, and Anti-Adhesive Activity of Sulfate Polysaccharide from against .Applications:Unspecified applicationReactive species:Unspecified reactive speciesBao Le,Duy Thanh Do,Hien Minh Nguyen,Bich Hang Do,Huong Thuy LePubMed 36433125
- Molecules (Basel, Switzerland) 27:2022Inhibition of NF-kB/IL-6/JAK2/STAT3 Pathway and Epithelial-Mesenchymal Transition in Breast Cancer Cells by Azilsartan.Applications:Unspecified applicationReactive species:Unspecified reactive speciesRania Alaaeldin,Fares E M Ali,Amany Abdlrehim Bekhit,Qing-Li Zhao,Moustafa FathyPubMed 36431925
Key facts
- Detection method
- Colorimetric
- Sample types
- Tissue Homogenate, Cell Lysate
- Assay type
- Semi-quantitative
- Reactive species
- Mouse, Human
- Range
- 6 - 600 µg/mL
- Assay time
- 1h 30m
- Sensitivity
- = 5 µg/mL
Reactivity data
| Application | Reactivity | Dilution info | Notes |
|---|---|---|---|
Application ELISA | Reactivity Reacts | Dilution info - | Notes - |
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Target data
Function
NF-kappa-B is a pleiotropic transcription factor present in almost all cell types and is the endpoint of a series of signal transduction events that are initiated by a vast array of stimuli related to many biological processes such as inflammation, immunity, differentiation, cell growth, tumorigenesis and apoptosis. NF-kappa-B is a homo- or heterodimeric complex formed by the Rel-like domain-containing proteins RELA/p65, RELB, NFKB1/p105, NFKB1/p50, REL and NFKB2/p52. The heterodimeric RELA-NFKB1 complex appears to be most abundant one. The dimers bind at kappa-B sites in the DNA of their target genes and the individual dimers have distinct preferences for different kappa-B sites that they can bind with distinguishable affinity and specificity. Different dimer combinations act as transcriptional activators or repressors, respectively. The NF-kappa-B heterodimeric RELA-NFKB1 and RELA-REL complexes, for instance, function as transcriptional activators. NF-kappa-B is controlled by various mechanisms of post-translational modification and subcellular compartmentalization as well as by interactions with other cofactors or corepressors. NF-kappa-B complexes are held in the cytoplasm in an inactive state complexed with members of the NF-kappa-B inhibitor (I-kappa-B) family. In a conventional activation pathway, I-kappa-B is phosphorylated by I-kappa-B kinases (IKKs) in response to different activators, subsequently degraded thus liberating the active NF-kappa-B complex which translocates to the nucleus. The inhibitory effect of I-kappa-B on NF-kappa-B through retention in the cytoplasm is exerted primarily through the interaction with RELA. RELA shows a weak DNA-binding site which could contribute directly to DNA binding in the NF-kappa-B complex. Beside its activity as a direct transcriptional activator, it is also able to modulate promoters accessibility to transcription factors and thereby indirectly regulate gene expression. Associates with chromatin at the NF-kappa-B promoter region via association with DDX1. Essential for cytokine gene expression in T-cells (PubMed:15790681). The NF-kappa-B homodimeric RELA-RELA complex appears to be involved in invasin-mediated activation of IL-8 expression. Key transcription factor regulating the IFN response during SARS-CoV-2 infection (PubMed:33440148).
Alternative names
NFKB3, RELA, Transcription factor p65, Nuclear factor NF-kappa-B p65 subunit, Nuclear factor of kappa light polypeptide gene enhancer in B-cells 3
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Recommended products
NF kappaB p65 ELISA Kit is a single-wash 90-min Simplestep used to quantify NF kappaB p65 with a sensitivity of 5 µg/mL. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.
- Colorimetric Sandwich ELISA - 450 nm readout : works on any standard plate reader
- Cited in over 30 citations
Key facts
- Detection method
- Colorimetric
- Sample types
- Tissue Homogenate, Cell Lysate
- Assay type
- Semi-quantitative
- Reactive species
- Mouse, Human
- Range
- 6 - 600 µg/mL
- Assay time
- 1h 30m
- Assay Platform
- Microplate
- Sensitivity
- = 5 µg/mL
Precision
Intra assay
| Sample | n | mean | SD | C.V. |
|---|---|---|---|---|
Sample HeLa Extract | n 6 | mean - | SD - | C.V. 2.8 |
Inter assay
| Sample | n | mean | SD | C.V. |
|---|---|---|---|---|
Sample HeLa Extract | n 3 | mean - | SD - | C.V. 8.7 |
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- +4°C
- Appropriate long-term storage conditions
- +4°C
- Storage information
- +4°C
Notes
Abcam's NFκB p65 in vitro SimpleStep ELISA™ (Enzyme-Linked Immunosorbent Assay) kit is designed for the semi-quantitative measurement of NFκB p65 protein in Human and mouse cells.
The SimpleStep ELISA™ employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.
As of October 2019, this kit was reformulated with new antibodies to maintain continued long term supply.
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
NF-kB p65 also known as RelA is a significant component of the NF-kB protein complex. This complex usually involves a molecular weight for p65 of approximately 65 kDa. NF-kB p65 is mechanistically a transcription factor that regulates genes involved in inflammation cell survival and immune response. Expression of p65 is widely seen in various cell types including immune cells and epithelial cells suggesting its role in numerous physiological processes.
- Biological function summary
NF-kB p65 acts as part of the larger NF-kB complex usually forming a heterodimer with other family members like p50. This complex translocates to the nucleus upon activation where it binds specific DNA sequences to regulate gene expression. The p65 subunit is essential for transactivating target genes involved in immune and inflammatory responses. Its regulation is important for proper cellular functioning especially in the maintenance of immune homeostasis and inflammatory response.
- Pathways
NF-kB p65 participates in critical pathways like the NF-kB signaling and Toll-like receptor pathways. These pathways are fundamental for initiating immune responses and contribute to the regulation of apoptosis and cellular stress responses. In the context of these pathways the IKK complex plays a pivotal role in NF-kB activation leading to the phosphorylation and subsequent degradation of inhibitors that retain NF-kB p65 in the cytoplasm therefore allowing its movement to the nucleus.
- Associated diseases and disorders
NF-kB p65 is implicated in conditions such as cancer and autoimmune diseases. Its dysregulation can lead to persistent activation of inflammatory pathways contributing to tumorigenesis and chronic inflammation. In the context of these diseases proteins such as the p50 subunit and the IKK complex are often involved reflecting the importance of tight regulation of NF-kB signaling in preventing pathological conditions.
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4 product images
Sandwich ELISA - NF kappaB p65 ELISA Kit (ab176648)
Typical cell lysate dilution series.
Example of a typical NFĸB p65 cell lysate dilution series. Background-subtracted data values (mean +/- SD) are graphed.
Sandwich ELISA - NF kappaB p65 ELISA Kit (ab176648)
Linearity of dilution in representative sample matrices. Cellular lysates were prepared at 3 concentrations in common media containing 1 x Cell Extraction Buffer PTR. Data from duplicate measurements of NFĸB p65 (Total) are normalized and plotted.
Sandwich ELISA - NF kappaB p65 ELISA Kit (ab176648)
NFĸB p65 phosphorylation in response to TNFα treatment.
Induction of NFĸB p65 (pS536) phosphorylation in MCF-7 cells in response to TNFα treatment. HeLa cells were cultured in 96-well tissue culture plates, and treated (10 min) with a dose-range of TNFα before cell lysis. Data from quadruplicate measurements of NFĸB p65 (pS536) are plotted and compared against total NFĸB p65 protein levels. Comparative NFĸB p65 (pS536) and NFĸB p65 (Total) data also shown by Western Blot.
Sandwich ELISA - NF kappaB p65 ELISA Kit (ab176648)
Comparison of NFĸB p65 expression in different cell lines.
Cell line analysis for Total NFĸB p65 from 200 μg/mL preparations of cell extracts. Data from triplicate measurements (mean +/- SD) are plotted and compared to 1X Cell Extraction Buffer PTR (zero).
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